Sf9昆虫细胞悬浮培养工艺的研究

对Sf9昆虫细胞在不同培养基、培养基中是否添加血清及不同生物反应器中的培养工艺进行了研究,发现Sf9细胞在Sf900Ⅱ无血清培养基中生长比在添加10％小牛血清的Grace培养基中更好,在Sf900Ⅱ无血清培养基14 L搅拌式生物反应器分批培养细胞密度可达1.4×107/mL.过程生化特性分析表明,总氨基酸的消耗及主要代谢副产物特别是乳酸及游离氨的积累是培养后期细胞密度降低及活性下降的重要原因.本研究为Sf9细胞生物反应器培养工艺优化及利用昆虫杆状病毒蛋白表达系统高效表达重组蛋白生产亚单位疫苗奠定了良好基础.

Study on Sf9 Insect Cell Suspension Culture Technology

Research on Sf9 insect cells suspension culture technology including with different medium,serum added or not and with different bioreactor were done.It found that Sf9 cells grew better in Sf900II serum-free medium than in Grace medium.The density of Sf9 cells in Sf900 II serum-free medium on 14 L stirred tank bioreactor reached 1.4×107/mL.The process of biochemical analysis showed that the total amino acid consumption and accumulation of metabolic byproduct such as lactic acid and ammonia were important factors for declining of cell density and viability during the late phase of cell culture.The study built a good foundation on optimizing bioreactor cell culture technology and efficiency expression of recombinant protein with baculovirus expression vector system for production of subunit vaccines.